Animal protein-free media for cultivation of cells

Abstract

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes.

What is claimed is: 
     
       1. An animal protein-free cell culture medium comprising putrescine and at least one protein hydrolysate, wherein the hydrolysate is derived from plants or yeast, and wherein the putrescine is present in the culture medium in a concentration ranging from about 0.5 to about 10 mg/L. 
     
     
       2. The animal protein-free cell culture medium of  claim 1 , wherein the medium further comprises cadaverine, spermidine, spermine, agmatine, or ornithine; or a combination thereof. 
     
     
       3. The animal protein-free cell culture medium of  claim 1 , wherein the protein hydrolysate is soy hydrolysate in a concentration ranging from about 0.05% (w/v) to about 5% (w/v). 
     
     
       4. The animal protein-free cell culture medium of  claim 1 , wherein the polyamine originates from a source other than a protein hydrolysate. 
     
     
       5. The animal protein-free cell culture medium of  claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to 30 mg/L. 
     
     
       6. The animal protein-free cell culture medium of  claim 1 , wherein the protein hydrolysate is present in the culture medium in a total concentration ranging from about 0.05% (w/v) to about 5% (w/v) for all protein hydrolysates. 
     
     
       7. The animal protein-free cell culture medium of  claim 1 , wherein the protein hydrolysate is derived from a plant selected from the group consisting of cereals and soy. 
     
     
       8. A method of cultivating cells, comprising the steps of:
 (a) providing an animal protein-free cell culture medium according to  claim 1 , and 
 (b) propagating the cells in the medium to form a cell culture. 
 
     
     
       9. The method of  claim 8 , wherein the cells are selected form the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells. 
     
     
       10. The method of  claim 8 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation. 
     
     
       11. A method for expressing a target protein, comprising the steps of:
 (a) providing a culture of cells that have been grown in an animal protein-free cell culture medium of  claim 1 ; 
 (b) introducing a nucleic acid sequence comprising a sequence coding for the target protein into the cells; 
 (c) selecting the cells carrying the nucleic acid sequence; and 
 (d) selectively inducing the expression of the target protein in the cells. 
 
     
     
       12. The method of  claim 11 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells. 
     
     
       13. The method of  claim 1 , wherein the cell/target protein combination is selected form the group consisting of CHO cells/coagulation factor VIII, BHK cells/erythropoietin, Epstein Barr virus transformed, and immortalized human B cells/human antibodies. 
     
     
       14. The method of  claim 11 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation. 
     
     
       15. A method for producing a virus, comprising the steps of:
 a) providing a culture of cells that have been grown in an animal protein-free cell culture medium of  claim 1 ; 
 (b) infecting the cells with the virus; 
 (c) selecting the virus-infected cells; and 
 (d) incubating the cells to propagate the virus. 
 
     
     
       16. The method of  claim 15 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells. 
     
     
       17. The method of  claim 15 , wherein the cell/virus combination is selected from the group consisting of Vero cell/attenuated vaccinia virus, Vero cell/vaccinia virus, Vero cell/hepatitis A virus, Vero cell/influenza virus, Vero cell/West Nile virus, Vero cell/SARS virus, and chicken embryo cells/FSME virus. 
     
     
       18. The method of  claim 15 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.